The complement system is a key component of a host's defense against microorganisms. However, some microbial pathogens have evolved to interact with and manipulate specific components of the complement cascade, tipping the balance against the host and in favor of the pathogen. This dynamic interplay between pathogens and components of the complement cascade presents challenges and opportunities for effective preventive and therapeutic strategies. Despite the importance, knowledge of the specific interactions that occur between the complement system and Bacillus anthracis is virtualy non-existent. Likewise, the biological consequences of such interactions remain unknown. Recent discoveries, primarily from our group, suggest that spores of B. anthracis have evolved a sophisticated mechanism to interact with the complement system. We propose a novel model in which the spore surface protein BclA directly interacts with complement component C1q and complement regulator factor H. Interaction with C1q mediates spore entry into different types of host cells in both a complement activation-dependent and activation-independent manner while interaction with factor H limits the extent of complement activation and promotes pathogen survival and persistence. If this model is correct, it will be significant in understanding the role of complement in the development of anthrax infections as well as providing a common mechanistic basis for; spore uptake by different types of host cells, the minimal inflammatory responses induced by spores and spore persistence in the lung, all of which are important features in the pathogenic process of B. anthracis. In addition, as BclA was shown to be protective in experimental animals, understanding the biological functions of BclA-complement interactions will have significant implications to future vaccine development. Furthermore, the studies proposed here are expected to have broad implications to complement-pathogen interactions in general. Consequently, two specific aims are proposed. In aim 1, we will determine the binding mechanisms of spore surface protein BclA to C1q and factor H, using a series of recombinant proteins as well as spores expressing different segments of the protein. In aim 2, we will determine the biological functions of spore interactions with C1q and factor H and the vaccine potential of recombinant BclA fragments. This will involve using mouse models deficient in specific complement components and spores isogenic for C1q binding or factor H binding. Mice will also be immunized with different fragments of BclA to determine their protective efficacy against both acute and persistent infections. The project will be carried out in collaboration with Dr. Rick Wetsel, Professor and Director, Research Center for Immunology and Autoimmune Diseases, Institute of Molecular Medicine, University of Texas Health Science Center (UTHSC), Houston, Texas. PUBLIC HEALTH RELEVANCE: The complement system is an essential component in host defense against microorganisms. The proposed project aims to investigate how spores of Bacillus anthracis, the causative agent for anthrax, actively engage components of the complement system and the biological consequences of such engagement. The project is likely to provide new and important insights into the molecular mechanisms underlying some of the key events in the establishment of infections and may lead to better vaccines to combat anthrax infections.